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The data on impact of chronic rhinosinusitis (CRS) on SARS-CoV-2 virus susceptibility and COVID-19 course were reviewed. CRS heterogeneity is determined by different types of inflammatory response. A heterogeneous CRS is divided into CRS without polyps and with nasal polyps (CRSwNP) is accounted for by diverse underlying immune responses. Hypersecretion of interleukins (IL)-4, IL-5, IL-13 in eosinophilic CRSwNP downmodulates angiotensin-converting enzyme (ACE)-2 receptor expression that should reduce SARS-CoV-2 infection risk because ACE2 is a main cellular tropism factor for SARS-CoV-2. In neutrophilic CRS type 1 immune response predominates, with activation of Th-1 cells, hypersecretion of interferon (IFN)-γ and tumor necrosis factor (TNF)-α to increase ACE2 expression. However, another data also show that hypoxemia level and pulmonary system damage did not differ between patients with CRS and CRS without polyps. Literature contradictions may be related to differences in availability of medical care, treatment of bronchial asthma (BA) as well as coverage of PCR testing. Regarding use of local or systemic glucocorticosteroids (GCS) effect on SARS-CoV-2 infection risk, some authors believe that GCS may increase COVID-19 severity and mortality, probably by downmodulating local innate immune response factors. According to other data, GCS may reduce ACE2 expression, or there is no relationship between previous GCS use, the incidence of COVID-19 and the frequency of treatment in the intensive care unit. Biological therapy of CRSwNP and BA with monoclonal antibodies did not aggravate COVID-19 severity and mortality risk. Although such data are currently limited, authors recommend not to interrupt such treatment during the epidemic, as well as continue taking leukotriene receptor blockers that can further inhibit major protease (Mpro) of the SARS-CoV-2 virus. However, according to international guidelines on COVID-19 treatment, using CRSwNP and asthma biological therapy should be discontinued until the patients recover completely. Allergen-specific immunotherapy (ASIT) should be interrupted in the case of confirmed COVID-19 due to a probability of developing severe COVID-19. After COVID-19, disturbances in the immune system may persist and possibly change the course of CRS, therefore requiring to modify therapeutic approaches for such patients. At the same time, the worldwide literature has been gradually accumulating information on pathogenesis underlying alterations in such patients including those with CRS, which requires development of new therapeutic approaches.

Acute COVID-19 is a viral infection caused by a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that results in dramatically decreased peripheral blood CD3+T cell count apparently due to alterations of thymic T cell maturation, that can persist long term afterwards. Therefore, we analyzed the levels of peripheral blood TRECs (T-cell receptor excision circles), and investigated the main alterations in peripheral blood T cell subsets in COVID-19 convalescents. We performed molecular quantification of TRECs with “TREC/KREC-AMP PS” kit and flow cytometric analysis of peripheral blood lymphocytes from three groups of patients. The first group contained 109 samples from COVID-19 convalescents (6–12 month post-acute COVID-19) with normal levels of TRECs (TRECn); the second was formed from COVID-19 convalescents (6–12 month post-acute COVID-19) with decreased levels of TRECs (TREClow, n = 29), and healthy control group (HC, n = 18). We noticed no significant differences between all three groups in CD3+T cell relative and absolute numbers. However, CD4+T cell frequencies were decreased in TREClow and TRECn groups compared to HC (40.8% (31.6; 50.1) and 46.4% (40.0; 53.0) vs 53.5% (47.36; 56.9), p 0.001 and р = 0.004, respectively). Furthermore, Th cell levels were decreased in TREClow patients vs HC and TRECn groups (701 cell/1 µL (478; 807) vs 1005 cell/1 µL (700; 1419), р = 0.020, and 876 cell/ 1 µL (661; 1046), р = 0.008, respectively). Finally, both groups of COVID-19 convalescents had increased frequencies of circulating CD8+T cells — 29.4% (20.7; 39.7) in TREClow group, 26.5% (21.1; 32.7) in TRECn group vs 21.3% (17.1; 26.0) in healthy controls (p = 0.024 and р = 0.026, respectively). In TRECn group, CD8+T cell count was elevated vs control range (508 cell/1 µL (372; 622) vs 356 cell/1 µL (247; 531), р = 0.044). Thus, COVID-19 convalescents (6–12 month post-acute COVID-19) showed an imbalance in CD4+and CD8+T cell level even at 6–12 months post-acute SARS-CoV-2 infection, and the observed changes in peripheral blood T cells could be closely related to the alterations in thymic T cell maturation and differentiation. Such a long-term decline in TREC levels in the circulation may have a profound impact on immune system functions and requires immunocorrection therapy.

The issues of recurrencies in chronic rhinosinusitis with nasal polyps (CRSwNP) still remain unresolved. Therefore, detection patients with uncontrolled clinical course of CRSwNP is required.The aim of the present study was to assess cytokine profile in nasal polyps as well as clinical characteristics of patients with CRSwNP at varying levels of therapeutic control.The study included 99 patients with chronic rhinosinusitis. The contents of interferon IFNγ, interleukin IL- 1β, IL-4, IL-5, IL-13, tumor necrosis factor TNFα, transforming growth factors TGF-β1, TGF-β2, TGF-β3 were measured in polyp tissue samples by means of multiplex analysis. The patients were treated according to a stepwise СRSwNP therapeutic algorithm [9]. Following observation for 5 years, all patients were divided into groups, as based on medical control degree. Group 1 included patients with mild CRSwNP, who mainly received stage I and II treatment for the entire observation period. Group 2 was presented by moderate- severity CRSwNP, with stage II or III therapy according to the referred algorithm. Group 3 included the patients with severe CRSwNP who received one or more stage IV courses. The patients underwent repeated SNOT-22 questionnaire, endoscopic examination, clinical assessment. In case of bronchial asthma (BA), ACQ-7 was repeated. Initial cytokine profile of nasal polyps was analyzed by the mentioned clinical groups.Results:After 5 years of observations in group 1 (mild CRSwNP), we found a minimal decrease in quality of life (SNOT-22), severity of nasal congestion, or smell impairment. Bronchial asthma duration in these patients was significantly lower compared to group 3, the patients had better asthma control level. Cytokine profile of nasal polyps was characterized by the highest IL-4 concentration, average values IL-1β, TNFα, IFNγ and minimal TGF-β1 values.In group 2 (moderate CRSwNP), we noted more pronouced impairment of smell, nasal congestion and quality of life. Bronchial asthma was less controlled than in group 1. The maximal concentrations of IFNγ, IL-1β, TNFα, IL-5, TGF-β1, TGF-β2 were registered.In group 3 with poor CRSwNP control, the highest SNOT-22 scores, severity of difficulty in nasal breathing, impaired sense of smell were revealed. Duration of bronchial asthma was longer, with lowest levels of medical control. In nasal polyps, minimal levels of IFNγ, IL-1β, TNFα, IL-4, IL-5, TGF-β2, TGF-β3 were noted.Treatment of patients depending on the clinical phenotypes of CRSwNP in the presence/absence of allergic rhinitis or bronchial asthma may improve control and reduce incidence of relapses in CRSwNP.

Objective: to evaluate the dynamics of the T- and B-cell immunity in patients with acute coronary syndrome (ACS) who have and have not had COVID-19, depending on the number of CD3+CD8+T-lymphocytes.Materials and methods: 65 men with ACS who underwent coronary artery stenting were examined. Immunological parameters were studied using flow cytometry, a complete blood count at baseline and 28 days after admission.Results: The maximum troponin level was observed in individuals with ACS who had recovered from COVID-19 and had a normal level of CD3+CD8+T cells. Stent thromboses and deaths occurred only among patients with a history of COVID-19, mainly with reduced CD3+CD8+T-cells, for which indicators of immune status were determined over time. The absolute numbers of T lymphocytes, T helper cells, late activated T lymphocytes, B lymphocytes (CD3-CD19+CD5+), B lymphocytes (CD45+CD3-CD19+) were minimal in individuals with low CD3+CD8+ T lymphocytes who had previously suffered from COVID-19, and significantly increased in their dynamics after 28 days. Natural killer cells significantly increased in dynamics in patients with initially low and normal CD3+CD8+T-lymphocytes who suffered from COVID-19.Conclusions: after stenting of the coronary arteries over time, in people with reduced CD3+CD8+Tlymphocytes and patients with COVID-19, T-lymphocytes (CD45+CD3+CD19-), T-helper cells, CD3+CD8+T-lymphocytes significantly increased, T-NK lymphocytes, NK lymphocytes, late-activated T-lymphocytes, T-regulatory lymphocytes and late-activated T-regulatory cells, B-lymphocytes, immunoglobulin G and complement fragment C3a decreased. T-regulatory lymphocytes and late-activated T-regulatory cells were significantly reduced in patients without prior COVID-19 with baseline low CD3+CD8+T-lymphocytes. In individuals with normal CD3+CD8+T-lymphocytes who recovered from COVID-19, T-lymphocytes (CD45+CD3+CD19-), NK-lymphocytes, and late-activated T-lymphocytes increased over time.

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has led to global morbidity and mortality. Some patients fully recover from COVID-19, whereas around 45% subjects suffer from various persistent symptoms (fatigue, cognitive impairment, impaired thermoregulation, skin diseases, etc.) for at least four months after SARS-CoV-2 infection regardless of disease severity. Such persistent post-infection effects are known as long-COVID, post-acute effects of COVID-19, or post-COVID state. SARS-CoV-2 infection is accompanied by damage to the innate immune system. Considering the role of natural killer cells and the activation of the complement system in COVID-19 as well as the regulatory properties related to CD46 and its potential involvement in cell virus entry, we found necessary to study immune system parameters associated with impairment of these innate immune cues on various leukocyte subpopulations in post-COVID patients. We studied 92 immune system parameters, including: pan-leukocyte markers for gated lymphocytes, phenotyping of T cells, T-helper inducers, cytotoxic T-lymphocytes, NK- and TNK-cells, T-regulatory cells/suppressors, B-lymphocytes, including B-memory cells, activated helpers, early activation of lymphocytes, activated T-lymphocytes, and late lymphocyte activation markers. Levels of total IgM, IgG, IgA, specific IgM, IgG to coronavirus COVID-19, C1-inhibitor, C3a, and C5a complement components were measured by enzyme immunoassay using Multiscan FC Thermoscientific enzyme immunoassay analyzer (China) and Vector Best reagent kits (Russia). A complete blood count was conducted to study 25 parameters: leukocyte, erythrocyte, and platelet hematopoietic lineages as well as the quantitative and qualitative composition of hematopoietic lineages. Our study results showed that in some patients, six months after suffering from COVID-19, there was a decrease in the level of NK cells (48%) and CD46+pan-leukocyte marker cells (64.5%). A decrease in NK cell levels was accompanied by increased level of total T- and B-lymphocytes, and altered platelet and erythroid hematopoietic lineages. In patients with reduced CD46 expression on T-lymphocytes, both their total count and NK cell count were significantly reduced. Our data also suggest that CD46 might be potentially involved in development of SARS-CoV-2 infection and the post-COVID state. Thus, in 50–65% of patients who have experienced SARS-CoV-2 infection, damage to the innate immune system persists after six months being accompanied by impaired erythroid and platelet hematopoietic lineages. The data obtained indicate a need for using immunocorrective therapy in such patients.

This study examines the long-term effects of SARS-CoV-2 infection on immune status. Given the prolonged and profound immune dysregulation observed during acute SARS-CoV-2 infection, it remains to be determined whether these changes translate into subsequent immune system dysfunction in recovering individuals. In this sense, the aim of the study was to study the parameters of the immune system in patients who had undergone SARS-CoV-2 infection. 150 patients who underwent SARS-CoV-2 infection were examined according to 96 parameters using flow cytometry. A complete blood count was performed using a Medonic device (Sweden); ELISA method determined the levels of general and specific IgM, IgG, IgA, compliment fragments (JSC Vector-Best, Russia). The activity of the phagocytes was studied according to the generally accepted method. The study found that at least four phenotypes of immune system disorders are detected in patients. The first two phenotypes are related to the impairment of innate immune system factors and are associated with a decrease in the number of CD46+ and NK cells. It has been observed that a decrease in CD46+ persists for a long time in a significant number of recovered patients, highlighted by the impaired expression of this marker in various subpopulations of lymphocytes. The decrease in the level of natural killers was accompanied by a compensatory increase in the number of T lymphocytes, mainly due to T helpers and TNK lymphocytes, and the growth of total memory B cells. Two other identified phenotypes are characterized by damage to acquired immune response factors and are associated with damage to B cells and T cytotoxic cells. The relationship of such disorders with damage to hematopoiesis erythrocyte and platelet sprouts, which contribute to the appearance of hypoxia and possible violation of the blood coagulation system, has been shown. Therefore, the results obtained indicate a long-term pronounced damage to the immune system in postCOVID patients that requires immunocorrection of these disorders.

The 2019 coronavirus disease (COVID-19) pandemic has had an unprecedented impact on health and economies around the world. Direct myocardial injury and cytokine storm, leading to destabilization of preexisting plaques and accelerated formation of new plaques, are two mechanisms that trigger the acute coronary syndrome in COVID-19. There is insufficient data on the immune status of patients with acute coronary syndrome who have undergone COVID-19. The aim of the study was to study T and B cell, humoral immunity depending on the number of cytotoxic T lymphocytes (CD8+) in patients with acute coronary syndrome who underwent COVID-19. Materials and methods of research: 65 men with unstable angina pectoris and acute myocardial infarction (acute coronary syndrome) from 40 to 65 years old, who had previously had COVID-19, were examined. A study of peripheral blood was carried out: complete blood count (Medonic device, Sweden), general and specific IgM, IgG, IgA, compliment fragments (Vector Best, Russia). Subpopulations of T and B lymphocytes were determined by flow cytometry. In persons with acute coronary syndrome who underwent COVID-19 with predominantly normal and elevated levels of cytotoxic T cells, a more severe course of the disease was observed: patients with acute myocardial infarction prevailed, they had longer mortality, longer treatment duration, and stent thrombosis was more common. In patients with elevated cytotoxic T cells, there was a maximum increase in erythrocytes, hemoglobin, hematocrit, lymphocytes of both the total number and subpopulations – T helpers, T-NK lymphocytes, NK lymphocytes, T lymphocytes of early and late activation, B1 and B2 lymphocytes, index of NBT-induced test. In patients with normal levels of NK cells, compared with other groups, there was an increase in spontaneous NBT activity and index, a significant decrease in C3a and C5a complement fragments. Prevalence of stent thrombosis and mortality in the group of patients with normal levels of cytotoxic T cells may indicate torpidity of the immune system in these patients with poor outcomes.

Introduction. Chronic rhinosinusitis with nasal polyps (CRSwNP) is considered a multifactorial disease. There are data on the contribution of fungi and viruses to the initiation and development of the inflammatory process, data on the effect of superantigens, biofilms and microbiota on the growth of polyps in the paranasal sinuses. Exacerbation of the disease in patients with CRSwNP leads to a significant decrease in the quality of life. Aim. To study the bacterial component of the microbiota of nasal and paranasal mucosa in patients with CRSwNP during remission and exacerbation. Materials and methods. 83 patients with CRSwNP were examined (44 patients in remission, 39 people in the period of exacerbation of the disease). A qualitative and quantitative analysis of bacterial component of the microbiota in all patients were carried out. Results. No significant differences in the qualitative and quantitative composition of the nasal cavity microbiota during exacerbation and remission of inflammatory process were observed, as well as before and after treatment of the CRSwNP exacerbation. The quantitative assessment of the identified microorganisms in the vast majority of cases was within the normal range.

Non-inflammatory exudative lesions of Reinke’ pos space present the most common cause of phonation disorders in representatives of vocal professions. The leading role in pathogenesis of this disorder is given to the nearly complete absence of lymphatic drainage of Reinke space and local deposition of tissue fluid. The mechanisms of its progression are of particular importance when determining the condition of mucosaassociated laryngeal tissue. The aim of our work was to evaluate the parameters of immunohistochemical lymphocyte typing in exudative lesions from the Reinke’s space. Materials and methods: The main group consisted of 40 patients, at the mean age of 43.2±2.1 years, exhibiting tumor-like exudative lesions of Reinke’s space. The biopsy material was taken from the vocal folds, including polyps, vocal nodules and Reinke’s edema. Myxoid and angiomatous types of the polyps were separately assessed. Videofibrolaryngoscopy was performed using an Olympus TYPE 150 bronchofibroscope (Germany). Morphological studies were carried out using a DMRXA microscope (Leika, Germany) by means of the ImageScopeM computer program (Germany). The uniformly treated sections were stained with Hematoxylin & Eosin (Biovitrum, Russia). Immunohistochemical quantitative assessment of the main T cell populations (CD3+, CD4+, CD8+), B cells (CD20+), histiocytes (CD68+), and the cells expressing bcl-2 and p53 cell regulators was carried out automatically using the BenchMarkXT immunohistotainer (Ventana, USA). The results were expressed as U/mm2. Results and Discussion. Some special features of cellular immunophenotype were revealed in exudative lesions of Reinke’s space. Reinke’s edema was characterized by high content of CD3+ lymphocytes, CD4+, CD8+, CD20+, p53 positive cells of the basal epithelium, as well as low numbers of CD68+, bcl-2 positive lymphocytes and cells of the basal epithelium. Myxoid type of polyps was characterized by low content of CD3+, CD4+, CD8+, CD20+ lymphocytes, bcl-2 positive lymphocytes and basal epithelium cells, CD68+ monocyte-macrophage cells, and high amounts of p53 positive basal epithelial cells. In the angioma-type polyps, we have registered low contents of CD3+, CD4+, CD8+, CD20+ lymphocytes, high numbers of monocyte-macrophage CD68+ cells, MMP-9+, bcl-2 positive lymphocytes, and low content of p53 positive cells of basal epithelium. The samples from the vocal nodules were characterized by low content of CD3+, CD4+, CD8+, CD20+ and p53 positive basal epithelial cells; high numbers of CD68+ cells (monocyte-macrophage series), MMP-9+ and bcl-2 positive lymphocytes.

Background. Leukotrienes play an important role in the pathogenesis of allergic rhinitis (AR) and eosinophilic type of chronic rhinosinusitis with nasal polyps (CRSwNP). There is a phenotype of CRSwNP in combination with AR, which has specifics of local inflammation.The aim of our study was to investigate the efficacy of using an antileukotriene drug in the treatment of AR in combination with CRSwNP.Materials and methods. 63 patients with AR and bilateral CRSwNP after endoscopic bilateral polypotomy were randomly divided into 2 groups. In the 1st group 32 people (age 50.28 ± 1.37 years) were prescribed a basic therapy with nasal spray of mometasone furoate at a daily dose of 400 µg in combination with montelukast 1 tab. 10 mg at night, in the 2nd group 31 people (age 50.31 ± 1, 16 years old) received only mometasone furoate monotherapy. Endoscopic examination of the nasal cavity was performed once every 3 months. The follow-up period was 1 year.Results. After 3 months in the 1st group of patients there was a recurrence of polyp growth was observed in 25% of cases, in the 2nd group in 35.5% of patients (p < 0.05). After 6 months, the number of relapses of CRSwNP decreased to 15.6% of cases in group 1 and to 22.6% in group 2 (p < 0.05). After 9 months in group 1 recurrence of NP was recorded in 12.5% of patients and nasal polyps were completely absent during endoscopic examination in 9.4% of cases, in the 2nd group, relapse was detected in 19.35% of patients (p < 0.05). 1 year after surgery, in group 1, relapse of NP was found in 12.5% of patients with AR and in 12.5% of cases was remission of the pathological process with cancellation of basic therapy. In group 2, recurrence of NP was in 16.1% of cases, there were no reasons for withdraw treatment of intranasal glucocorticosteroids in this group.Discussion. The clinical effectiveness of the addition of Montelukast to basic therapy has been reflected in a reduction in the growth rate of polyposic vegetation, the number of repeated operations and the stabilization of the flow of chronic inflammatory process.Conclusions. In the case of the clinical phenotype of AR with CRSwNP, the addition of a leukotriene receptor blocker montelukast to the basic therapy of intranasal glucocorticosteroids made it possible to improve drug control of both diseases and reduce the frequency of CRSwNP relapses.

Chronic rhinosinusitis with nasal polyps (CRSP) is a heterogenous disease. We have earlier detected differences in severity of clinical manifestations, cellular infiltration degree shown in nasal polyps, of eosinophil-to-neutrophil ratio, efficacy of intranasal glucocorticosteroid baseline therapy, and various inflammatory patterns for several cytokines on the mRNA expression level in different phenotypes with isolated CRSP cases, CRSP associated with respiratory allergy (RA), or non-allergic bronchial asthma.The purpose of this work was to study the cytokines of TGF-â family in the tissues of nasal polyps in patients with different CRSP phenotypes. The research involved 292 patients suffering from CRSP divided into 3 phenotypic groups. Group I consisted of patients with isolated CRSP free of associated BA and/or sensitization to atopic allergens. Group II included patients with CRSP combined with respiratory allergy and was further divided into two subgroups. I.e., Group 2a comprised patients with CRSP, allergic BA (aBA), and allergic rhinitis (AR), while the patients with CRSP, AR, and non-allergic BA were placed to the group 2b. The patients suffering from CRSP complicated with non-allergic BA were allocated to the group III. The patients with hypertrophic rhinitis served as control. The levels of TGF-â1, TGF-â2, and TGF-â3 proteins (pg/mg) were measured by means of multiplex immunoassay approach in supernates of tissue homogenates from nasal polyps removed by surgery, and in posterior parts of inferior nasal conchae. The total protein level was determined in tissue supernatant, with cytokine contents recalculated for the mg/ml protein concentration for standardization of measurements.In the control group, trace concentrations of all three growth factors were detected. Significant difference in protein contents was found for the studied cytokines, depending on CRSP phenotype. The levels of TGF-â1 and TGF-â2 were statistically lower in isolated CRSP than in other groups of comorbid CRSP patients. TGF-â1 and TGF-â2 concentrations were significantly lower in CRSP + allergic BA group IIa than in CRSP + nonallergic BA and CRSP + RA groups. The amount of TGF-â3 cytokine was maximal in CRSP + non-allergic BA group III compared to the patients with isolated CRSP of group I and CRSP + non-allergic BA group 2a.Conclusions.The high level of all three TGF-â isoforms in patients with CRSP compared to the control group suggested a high potential of mucous membranes of paranasal sinuses for active tissue remodeling followed by nasal polype formation.Different mechanisms were presumed for development of local pathological process in different clinical phenotypes of CRSP, depending on the comorbid pathology, especially, BA or respiratory disorders.Minimal TGF-â1 and TGF-â2 levels were detected in isolated CRSP.The highest concentrations of TGF-â1, TGF-â2, and TGF-â3 were discovered in the patients with CRSP accompanied by non-allergic BA as compared to the groups with isolated CRSP and CRSP+allergic BA.5. Determination of TGF-â1, TGF-â2, and TGF-â3 levels can serve as an additional criterion for differentiating between the mechanisms of mucous membrane damage in local pathological process in tissues of comorbid patients with different CRSP phenotypes.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) belongs to the group of growth cytokines (hematopoietins) that regulate proliferation and differentiation of myeloid lineage cells. Recently, a lot of new data have accumulated, indicating the presence of a number of previously unknown biological effects in GM-CSF and synthetic peptides of its active center, which open up new prospects for their wide clinical use.The review outlines current understanding of the structure, functions, and mechanisms of GM-CSF action and concerns the structure of its receptor. The GM-CSF producer cells are characterized, as well as target cells (effector cells) responding to this cytokine are also presented. The known mechanisms of intracellular signaling involved into the GM-CSF/receptor interaction are described. The main pleiotropic effects of this cytokine as a factor of hematopoiesis and an immunostimulating agent are characterized. The previously known and recently found immunobiological effects of this cytokine, its recombinant forms and synthetic analogues of its active center are discussed.Participation of GM-CSF in hematopoiesis and differentiation of myeloid cells, the effects of this cytokine on the functional activity of immunocompetent populations (lymphocytes, macrophages, neutrophils, dendritic cells) and tissue cells were characterized. The influence of GM-CSF on the development and course of infectious and inflammatory processes, its role in the creation of combined vaccines is reviewed. Clinical data on usage of GM-CSF and its recombinant forms in hematology, immunology, oncology, reproductive medicine and in the treatment of systemic autoimmune processes and infectious diseases are presented.The recently discovered immunobiological properties of synthetic peptides derived from active center of GM-CSF are summarized, indicating that they exhibit immunotropic and hematopoietic effects, as well as antimicrobial activity against Gram-negative and Gram-positive bacteria, viruses, and tissue repair (effect on the rate of wound healing), which is not typical to the whole GM-CSF molecule. We discuss the prospects for clinical applications of synthetic GM-CSF analogue (ZP2 peptide), and an opportunity of creating new cosmetics and pharmaceuticals with combined immunostimulating, antimicrobial and reparative properties on its basis.The review expands the view on potential usage of cytokine therapy in the treatment of various infectious and non-infectious diseases in humans, and is addressing a wide range of specialists working in the field of allergology and immunology, infectology and regenerative medicine.

Treatment of chronic viral infections accompanied by permanent virus persistence in the target epitopes of the oral cavity, skin, urogenital tract is complicated by virtual lack of available drugs exerting combined systemic virulicidal and immunomodulatory effects. Here we demonstrate clinical and immunological efficacy of combined therapy in treatment of Epstein–Barr virus (EBV)-associated chronic infections. The aim of the study was to evaluate the clinical and immunological efficacy of combined etiopathogenetic therapy using the Acegram cosmetic product in patients with EBV-associated chronic infections. Materials and methods. There were enrolled 40 patients monitored before treatment as well as 20 patients followed up after combination therapy (cycle therapy consisted of oral valaciclovir (Valtrex) applied at dose of 500 μg twice a day for 10 days, glucosaminylmuramyldipeptide (Licopid) — 10 mg 2 twice a day for 10 days, topical irrigation for mucous membranes with granulocyte-macrophage colony-stimulating factor active center-derived peptide (Acegram-spray) 3 times a day for 10 days. If necessary, treatment courses were repeated 20 days after the onset. All patients were examined for the presence of EBV genomes in the oral fluid and blood using the qualitative and quantitative polymerase chain reaction (PCR) using the DNA technology test system (Russia) on a DT-Lite device prior treatment and 30, 60 days post-therapy time points. In addition, serum samples were analyzed for level of class G immunoglobulins specific to the EBV nuclear and capsid antigens by using enzyme immunoassay (test systems manufactured by CJSC Vector Best, Russia) as well as immune status (clinical methods, enu flow cytometry evaluation of the phagocytic activity of neutrophils, ELISA method). Results. Use of single or two course combination therapy in subjects with fully eradicated EBV carriage associated with reversed clinical symptoms was accompanied by recovered immune system status (T and B cells, T-helper cells, CD3+ CD25+ cells, phagocytosis parameters). A non-invasive approach proposed for controlling virus elimination in the oral fluid by using polymerase chain reaction method may serve as to objectively monitor therapeutic efficacy.

It is known that the immunological component of nearly any dysregulatory pathology is manifested by the development of new functional systems providing adaptation reactions, as well as by the presence of allostasis, reflecting other forms that are different from the normal parameters, and variants of mutually supporting relationships that occur at different structural and functional levels of the body. A long stay in the areas of military campaigns inevitably has a negative impact on the re-adaptation for civilian life, and it is accompanied by occurence of pathological functional connections that lead to dysregulation of the basic functions, including those of nervous and immune systems. In this regard, we studied the features of immune cell functioning in the patients with early forms of chronic cerebral ischemia accompanied by hyperlipoproteinemia, and we have made a comprehensive assessment of the factors characterizing morphological and functional state of endothelium. A representative survey included 87 patients who participated in the Afghan campaign with early forms of cerebral circulatory failure. The subjects exhibited objective and subjective signs of underlying vascular disease, typical to the earliest manifestations of chronic cerebrovascular insufficiency (CVVN) manifesting as headache, dizziness, noise in the head, memory impairment, decreased performance. All the patients underwent a comprehensive examination, i.e., expanded lipidogram, determination of vasoactive factors in the blood and cultural fluid, the number of circulating desquamated endotheliocytes, thus reflecting morphological signs of endothelial damage, additional ultrasound Doppler sonography of the main vessels, transcranial Doppler and electroencephalography were carried out.It is known that endothelial cells are independently capable of modulating vascular tone, depending on the current clinical condition, and it may be also subjected to remodeling rearrangements with involvement into the pathological process, e.g., in atheromatosis. In this regard, endothelial dysfunction is a key factor in developing vascular pathology, affecting the systemic level. According to results of the study, it has been established that the main progression mechanism for the early forms of CVVN is the development of endothelial dysfunction. The condition has multifactorial genesis in which immune mechanisms are directly involved. Hyperlipoproteinemia established in more than 90% of cases, is one of the most important metabolic triggering factors for development of immune reactions, which is also promoted by imbalance of vasoform and thrombogenic factors, by presence of oxidative and nitrosative cellular stress markers which generally contribute to increase in cerebral vascular remodeling and the basis for the formation of organic pathology of the central nervous system.

Antibacterial drugs are routinely used in therapy of periodontal diseases. However, increasing incidence of antibiotics resistance necessitates development of novel therapeutic approaches for oral diseases. Currently, newly designed antibacterial agents based on natural, semi-synthetic and synthetic peptides is the most promising approach in dentistry. Among them is a product containing synthetic peptide (ZP2) replicating active site of granulocyte-macrophage colonystimulating factor (GM-CSF) as the main active ingredient in Atsegram-spray (manufactured by Academic Innovation Research Center, Chelyabinsk). Our study was aimed at assessing efficacy of a combination therapy of inflammatory periodontal diseases by introducing Atsegram-spray as well as examining potential relationship between peptide ZP2-related biological properties, physicochemical properties of the spray and mechanism of antibacterial and immunotropic action for substantiating its application on oral mucosa. During the first stage experiments, it was found that the peptide ZP2 was able to trigger lymphocyte blast transformation in vitro indicating that it might influence cell proliferation and exhibit marked immunotropic activity. Next, we assessed potential effects of the peptide ZP2 on biofilm formation mediated by staphylococcal clinical isolates. It was shown that peptide ZP2 inhibited biofilm formation in 75.0±9.0% of S. aureus and 50.0±15.1% of S. epidermidis strains, with mean inhibition index of biofilm formation reaching 25.1±3.8 and 50.4±6.0%, respectively. However, peptide ZP2 in 8.3–25.0% of staphylococcal clinical isolates was found to stimulate/lack effect on biofilm formation by 14.9–48.5 and 16.7–25.0% cultures, respectively. Thus, the synthetic peptide ZP2 exerts divergent, but mainly inhibitory effects on biofilm formation with staphylococcal clinical strains, which are characterized by inter- and intraspecific (strain) variability. Use of a synthetic peptide-based spray in antibacterial therapy of mild chronic generalized periodontitis (main group) one month after the onset was found to improve oral hygiene by 28.5% as well as decrease PMA index and gum bleeding index by 82.8 and 100%, respectively. In contrast, such parameters in patients receiving basic therapy (comparison group) were lower on average by 2-fold. While analyzing physicochemical properties of the spray such as pH, buffer capacity and solution osmotic pressure, it was found that they were related to the antibacterial mechanism of drug activity and efficacy in treatment of inflammatory periodontal diseases. Thus, assessing peptide ZP2-related biological properties and physicochemical parameters of the spray allows to evaluate their role in mechanism of previously unknown antibacterial and immunotropic activity. These findings confirm feasibility and efficacy of using Atsegram-spray in dentistry as an alternative means to antimicrobial agents, such as antibacterial drugs.

Despite numerous attempts to control the course of chronic rhinosinusitis with nasal polyps (CRSwNP) by means of pharmacological treatment and new surgical approaches, the majority of patients experience lifelong persistence of this disorder, at recurrence rates of 50-60% within 18 months after surgical treatment. Since CRSwNP is a chronic persistent inflammatory process, it affects the entire body condition, including the state of systemic immune response. An elevation of NK (CD3-CD16+CD56+), activated NK (CD8+CD3-), NKT cells (CD16+CD56+CD3+), Treg (CD4+CD25brightCD127low to neg) cells and activated T-lymphocytes (CD3+CD25+) was revealed elsewhere among all the patients with CRSwNP, using a flow ytometry method. There was no difference between various disease phenotypes. We analyzed the status of cellular component of systemic immunity, dependent on clinical course of the disease and efficiency of the administered therapy of CRSwNP. The patients were divided into three subgroups. The follow-up period was 1 year. The first group comprised the patients who showed positive dynamics after conservative therapy, resulting into regression of nasal polyps and their grade than a year ago. The second group included the patients in whom the size of polyps remained the same. The third group included the patients with higher incidence of nasal polyps than a year ago.We have shown a decrease of Treg, NKT cells, NK and activated NK, cytotoxic T-lymphocytes (CD3+CD8+), activated T-cell numbers in clinical group 3 with aggressive growth of polyps and low effect of standard therapy, which may cause deterioration of the immune system cellular populations, accompanied by presence of persistent productive inflammatory process of nasal cavity and paranasal sinuses. In the second group, a significant elevation of total lymphocyte number, total and activated T cells, T helpers (CD3+CD4+), cytotoxic T lymphocytes, NK and NKT cells was shown. Meanwhile, a decrease in absolute number of activated NK was observed despite the NK growth. Therefore, we can assume that the mechanism of their activation was disturbed and compensated by production of NKT cells and cytotoxic T lymphocytes. Moreover, we have shown in this group that the absolute number of Treg cells is increased; and these cells had a suppressive influence on effector cells of adaptive immune response, thus inducing incomplete elimination of infectious agents, which contribute to permanent incomplete course of inflammatory process. Chronic inflammatory process in CRSwNP affects systemic cellular immunity depending on the morbidity characteristics in the course of pathological process. The maximal intensity of systemic cellular immunity is observed in the group of patients that require permanent basic drug therapy. In case of aggressive CRSwNP and failure of standard drug therapy, we observed a decrease in absolute numbers of effector cells, along with decreased Treg lymphocyte numbers which may explain inefficient immune regulation of inflammatory process and medical interventions in this group of patients.

Purpose: to study the effectiveness of aflibercept in the treatment of neovascular age-related macular degeneration (AMD) combined with cataract phacoemulsification with intraocular lens (IOL) implantation. Material and methods. A short-term, prospective, controlled, non randomized comparative study of the effect of aflibercept on the changes of visual acuity, the morphometric parameters of the macular area of the retina, and the immunological ophthalmologic status was carried out on 31 patients with neovascular AMD within 1 month after medication usage start. Results. Intravitreal injection of aflibercept leads to a decrease in diffuse retinal edema in the macular area, the size of neuro- and pigment epithelium detachment, and the size of choroidal neovascularization, which in a number of cases was accompanied by an increase in visual acuity, especially with a simultaneous surgical treatment of cataract. Phacoemulsification with IOL implantation neither deteriorates the course of AMD nor affects aflibercept efficacy. The determination of the immune status of tears in the early period after intravitreal administration of aflibercept could not be seen as a relevant method of drug effectiveness assessment. Conclusion. Intravitreal administration of vascular endothelial growth factor inhibitor, aflibercept (Eilea, Bayer Pharma AG, Germany) is an effective method of neovascular AMD treatment including the cases of simultaneous cataract surgery. For citations: Kuznetsov A.A., Tur E.V., Zurochka A.V., Rykun V.S. Efficacy of aflibercept in patients with neovascular age-related macular degeneration when combined with cataract surgery Russian ophthalmological journal. 2017; 10 (4): 20-8. doi: 10.21516/2072-0076-2017-10-4-20-28 (in Russian).

A non-specific stress-induced overload of limbic-reticular emotiogenic structures under the deficiency conditions of endogenous energy resources is the main mechanism for development of asthenic disorders with early developing chronic brain ischemia among veterans of recent wars, upon their return to civilian life. We observed a group of 30 Afghan veterans with early forms of chronic brain ischemia and manifestations of psychogenic asthenic syndrome. When treating psychogenic asthenic syndrome in the veterans of recent military conflicts, an adamantane derivative administered as a course treatment, at a dose of 50 mg 2 times daily causes psychostimulant, anxiolytic, vegetotropic, antihypotymic, and hypnotic pharmacological effects, with reduction of main psychopathological and somatoneurological symptoms.A notable immunotropic effect of the drug was revealed, being expressed as enhanced immunopoiesis; activation of innate defense mechanisms; expansion of Treg lymphocyte population, suggestive for increased suppressor effect upon the mechanisms of autoimmune aggression; implementation of cell differentiation/ proliferative effect; anti-apoptogenic properties; anti-inflammatory activity (reduction of TNFα, IL-8, hsCRP, along with IL-2 increase) associated with reduced functional activity of mononuclear cells; vasotropic action expressed as normalized balance of vasoactive factors (nitric oxide, endothelin-1), decrease of nitrotyrosine concentration, suggesting the nitrosylation stress reduction. The actoprotective drug effect is implemented due to a combined psychostimulant, vasotropic, proliferative effects. It may be expressed in terms of higher efficiency and optimization of systemic hemodynamic parameters.

We studied clinical efficacy of sublingual immunotherapy in children with combined respiratory forms of pollen allergy before and after three courses of treatment. The levels of G-CSF, GM-CSF, IL-10, IL-12p70, IFNγ, IL-13, IL-17A, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, TNFα, IL-8, MCP-1, MIP-1β were measured in nasal secretions. In remission state, the children with combined forms of respiratory allergy to pollen allergens showed local signs of chronic inflammatory process (in the nasal mucosa) associated with disturbed immune response, with increased synthesis of proinflammatory cytokines and decreased production anti-inflammatory cytokines, as well as enhanced reactions aimed for allergen elimination. Following clinical success of sublingual immunotherapy, we have shown a reduction in immunological imbalance, due to decreased activity of the systems modulating severity of inflammation and production of pro-inflammatory cytokines.

Urban ecological problems are associated with industrial pollution emissions, waste of metallurgical production, unhealthy lifestyle and reproductive behavior lead to a steady rise in cancer pathology in theChelyabinskregion. Women in early reproductive age living in areas of ecological concern with suspected mastopathies, exhibit significantly higher levels of tumor markers associated with the tumor cell membranes of (CA19-9, SA15-5, CA125), CEA (carcinoembryonic antigen). Therefore, it seems necessary to perform massive laboratory programs for oncological screening, especially, among young working population, in order to identify breast diseases, and to arrange appropriate risk groups at this basis, aiming for development of effective preventive measures for the future.

Formation of the earliest forms of chronic brain ischemia veterans of modern wars accompanied by an increase in the systemic circulation of the population of T lymphocytes and monocytes, reflecting the activation of central mechanisms lymphopoiesis. In step vascular encephalopathy is an increase in circulating pool of T lymphocytes expressing the activation markers early positive reflecting readiness cells to IL-2 dependent proliferation. When progessirovanii chronic brain ischemia decreased levels of circulating T-regulatory cells, which may reflect a violation of self-tolerance in relation to brain antigens.

Patterns of immune response, cytokine- and NO-ergic mechanisms and their role in the development of basal-cell skin cancer were evaluated in population of an industrialized region. Multiple forms of basal-cell skin cancer and primary multiple cancer were characterized by the evidence of Th2-dependent imbalance of immune response, decreased cytolytic potential, increase of EGF, TGF-β, IL-1β, IL-4, IFNγ, lactoferrin levels, thus suggesting a particular immune defense strategy in malignant growth.

Abstract. Several endogenic and synthetic peptides were investigated in order to reveal their antibacterial activity. The study of dose-depended effects of antibacterial activity of the immunotropic peptide ZP-2, which is included in GM-CSF active center, shown significant antibacterial activity of ZP-2 by action range and by rate of bactericidal activity in case of low concentration. Combined application of substance derived from supernatant of CD34+CD45-cells and synthetic peptide ZP-2 enhances significantly actions of each other. Probably, this effect is connected with various mechanisms of their influence on bacteria. The obtained facts indicate that synthetic peptide has not only immunostimulating action but it also promotes a direct antibacterial activity. These data support an idea that synthetic peptides have good perspectives in case of development new antibacterial drugs with unique characteristics.

Incidence of different types of dislipoproteinemias (DLPs) and a balance between apoptosis andproliferation of lymphocytes were studied in males exhibiting initial manifestations of cerebral blood flowinsufficiency (ICBI). In general, high intensity of lymphocyte activation was revealed in patients with ICBI,with amplified proliferative potential demonstrable in this group. We have also shown a trend to increasednumbers of T and B lymphocytes expressing antiapoptogenic bcl-2 protein. The changes in apoptosis and cellproliferation intensity were more pronounced in IIA and IIB types of DLP. In these groups, unidirectionalchanges of FasR expression were observed. However, the parameters reflecting readiness to apoptosis werechanged multidirectionally. Cholesterol and oxysterols transported to the cells under excessive dyslipidemiamay influence activities of transcription factor genes (bcl-2, Fas-regulatory gene (TDAG51), thus, probably,causing an imbalance between cellular proliferation and apoptosis.

Abstract. T-lymphocytes play an important role in elimination of tumor cells, in reactions of a transplant against graft and graft versus host disease, in slow-type hypersensitivity, and other reactions directed for maintenance of homeostasis. Along with CD3, an antigen-specific T-cellular receptor (TCR) is another common marker of T-cells. There are two types of TcR – αβ-TcR and γδ-TcR that differ in ontogenetic and functional properties. γδ-T-cells play a significant role in protection of organism against various types of infections, and determination of their amounts should be an integral part of the analysis of patients’ immune status. To these purposes, a multi-colour analysis shuld be used, applying the following combinations of monoclonal antibodies: CD3/CD4/CD8/CD45 and αβ-TcR/γδ-TcR/CD3/CD45. Multi-colour staining and multi-step gating allow of carrying out multiparametric analysis of peripheral blood with high accuracy and reliability. The proposed approach considerably facilitates interpretation of results obtained, and it allows of judging about immune system functioning in various pathological conditions.

Abstract. The purpose of this article was to make a broad audience of experts familiar to new developments in the method of flow cytometry that have found applications in medical and biologic studies, in order of their further implementation into everyday practice. These applications include the following approcahes: detection of antigen-specific T-cells by using of tetramer technology, flow cytometric determination of cytokines in biological liquids, determination of in vitro sensitivity of basophilic granulocytes to allergen effects, detection of Th1 and Т-reg cells by their cell surface markers. When using multiparametric analysis, a multi-step gating and other new technologies, the flow cytometry technique allows of location and tracing the majority of processes occurring in development of immune response. When studying these dynamic events, we get an opportunity to react adequately to apropriate changes, and to develop new approaches to correct altered cellular activities, that they should determinate.

Abstract. The article considers an original approach to individual dose selection and efficiency evaluation of immunity-targeted drugs applied for treatment of children with trauma, based on examination of phagocyte locomotive functions.

Abstract. A study of leukocyte regulation system in patients with exacerbated bronchial asthma (BA) was performed by means of measuring spontaneous and induced production of certain cytokines (IL-4, IL-1b, IL-10 and IFNg) in whole blood, and drawing some specific coefficients reflecting production of the cytokines, as compared to control values. Group 1 consisted of mixed-type BA patients, with mild clinical manifestations at exacerbation stage. A control group included twelve healthy persons. The cytokine network was evaluated according to spontaneous production of cytokines (IL-4, IL-1b, IL-10 and IFNg), or their induction with standard mitogens (PHA and E. coli LPS). The results obtained have shown that in BA as a clinical model of chronic inflammation, synthesis and activation of the cytokines depend on the phase of disease. At stage of exacerbation, an imbalance occurs between pro- and anti-inflammatory cytokines, with predominance of anti-inflammatory IL-10, which is targeted to suppression of bronchial inflammation.

Abstract. We analyzed efficiency of application for analytic control parameters, i.e., «T-cell replicates» and «T-cell summary» in a sample of 174 HIV-infected patients, independently of age or disease stage. The patients were divided in two groups. Group 1 consisted of patients with total CD45+CD3+CD4+ and CD45+CD3+CD8+ lymphocyte concentrations being similar to relative and absolute concentrations of CD45+CD3+ lymphocytes, or the difference between absolute measurements not exceeding «T-cell replicates» differences. Group 1 included 36.78% of the whole cohort. Group 2 consisted of patients with total CD45+CD3+CD4+ & CD45+CD3+CD8+ lymphocytes concentrations being lower than relative and absolute concentrations of CD45+CD3+ lymphocytes, and the differences in absolute values being higher than «T-cell replicates» differences (41.38% of total sample). Such difference between control sums may be connected with high concentrations of double-negative γδ-T-cells (CD45+CD3+CD4–CD8–) in the specimens under study. (Med. Immunol., 2008, vol. 10, N 4-5, pp 463-466).

Abstract. Temporary immunodeficiency is often associated with various pathological conditions, such as myocardial infarction, severe trauma, or major surgery. However, there are no clinical criterions indicative for presence or absence of such immune deficiency. Meanwhile, clinical signs of infectious complications may be absent because of deficient immune response.A technique for monitoring septic conditions in the patients with open traumas and during postsurgical period has been proposed, employing a flow-cytometric approach. The evaluation principle for septic conditions consists of measuring expression of HLA-DR antigens at the surface of peripheral blood monocytes.As a criterion of patient evaluation in septic state, a relative amount of monocytes expressing HLA-DR may be applied, and the disease prognosis is considered as favorable, if the amounts of positive cells exceed 40% by day 5 after the patient was admitted to the hospital, and an adequate treatment was carried out. This technique may find wide application for estimation and monitoring of septic conditions in the patients. (Med. Immunol., vol. 10, N 4-5, pp 379-388).

Abstract. Determination of lymphocyte subpopulations and their phenotypes is an important diagnostic feature, in order to elucidate some disturbances connected with immune system functioning. However, insufficient data are obtained when analyzing only major populations of peripheral lymphocytes. In order to perform clinical diagnostics, the data about minor lymphocytic populations and activated cellular pools seem to be more pertinent.Studies of peripheral blood cell subpopulations of healthy donors performed in different Russian regions allowed to assess quantitative distribution intervals for both major and minor immune cell subpopulations in humans. The results obtained, as compared with data from literature, provide an evidence for similar reference intervals for main immune cell subpopulations in healthy donors, independent on their habitation area.Present work has resulted into development of algorithms for cytometric studies and generation of certain panels of monoclonal antibodies enabling evaluation of all main lymphocyte subpopulations, as well as their minor subsets participating in emerging immune response. The distribution intervals have been estimated for such minor subpopulations, as B1- and B2-lymphocytes, memory B-cells, γδ- and αβT-cells, regulatory and naїve T-cells, cytotoxic and secretory NK-cell polupations.The results of present study, while been performed with peripheral blood of healthy donors, may provide a basis of reference values when studying subpopulation profile of immune cells.

Abstract. Flow cytometry is considered as a modern technology for fast measurements of cellular characteristics, their organelles, and processes occurring within them. It is regarded as an efficient solution in many important areas of cell biology, immunology and cellular engineering. Present article bears on main developments in flow cytometry and their applications in medical and biological practice. Usage of modern achievements in fluorescent dyes, progress in laser and computer technologies, as well as potent software, resulted in wide application of this technique in medical practice. Accordingly, usage of monoclonal antibodies conjugated to different fluorochromes has led to elaboration of multiparametric analysis and did sufficiently simplify specialized works aimed for diagnostics of various immune disorders. The new directions in flow cytometry, e.g., flow cytoenzymology, provide wide opportunities for detailed identification of damaged or altered cells, and taking adequate decisions in treatment of detected pathological changes. The authors suggest that this article could initiate a series of publications concerning usage of this technology and its modern applications in broad laboratory practice.

Abstract. This work was aimed for studying immunity markers, especially, T-regulatory cells, in patients suffering from polypous rhinosinusitis and treated with nasal steroids, in frame of integrated preventive treatment. The study included fifteen patients of both sexes (18 to 60 years of age), and eighteen persons with difficulties of nasal breathing not associated with inflammatory events. T-regulatory cells were determined by a specific set of membrane receptors (CD4+, CD25+, CD127-). We have shown a significant reduction in T-regulatory cell contents prior to treatment, followed by their increase to normal levels in the course of therapy.

Abstract. A study of leukocyte regulation system in a remission stage of bronchial asthma (BA) patients included following tests: CD-typing of lymphocytes, investigations in humoral and phagocytic mechanisms of immunity, evaluation of bronchoalveolar lavage characteristics, measurements of spontaneous production of IL-4, IL-1β, IL-10 and IFNγ cytokines in whole blood supernates and bronchoalveolar lavage, as compared with control samples. The first group represented mild-stage, mixed-type BA patients (both allergy- and infection-dependent), being in remission state. The second (control) group consisted of sixteen conditionally healthy patients.The results have shown that, even at early stages of BA development, some components of chronic inflammatory process are formed. They could be characterized as accumulation of different inflammatory cells, i.e., eosinophils, neutrophils, alveolar macrophages and monocytes at the local level. In parallel, some changes in T-, B-subpopulations and phagocytic compartment occur at systemic level. All these events comprise a basis for changes in type and quantity of the cytokines produced. It was established that the profile of cytokine secretion in peripheral blood is identical to the cytokine secretion profile in lung tissue.

We have performed immunophenotyping of lymphocytes in 150 HIV-positive patients, by means of flow cytometry. All the persons under study were classified into three groups, depending on absolute contents of CD3+CD4+T-lymphocytes in peripheral blood, according to HIV infection staging in adults and adolescents, as amended by CDC (USA). It was revealed that, at each subsequent stage of disease, a decrease in absolute numbers of T-helpers is accompanied by drop in relative and absolute numbers of lymphocytes, as well as in total T-lymphocyte numbers, and decreased amounts of absolute cytotoxic T-lymphocytes levels, combined with increase in their relative contents, along with decreased CD3+CD4+/CD3+CD8+ ratio, diminished levels of double-positive subpopulation, decrease in relative and absolute levels of αβТ-lymphocytes and significant drop in absolute levels of γδТ-cells, while their relative amounts remained normal. These findings demonstrate a pattern typical to the absence of immunological reactions in response to antigens persisting in the organism.

Abstract. We have performed immunophenotyping of lymphocytes, employing a flow cytometry approach, along with evaluation of spontaneous or mitogen-induced production of cytokines (IFNγ, IL-4, IL-1β, IL - 10), in the patients with atopic and bacterial (mixed-type) bronchial asthma. We have detected mostly similar changes in lymphocyte parameters. However, among the patients with bacterial asthma, an imbalance of immune cells was more pronounced. Some common alterations of cytokine production were found, i.e., a decrease in IFNγ and IL-1β production, along with increased IL-4 and IL-10. These changes were more significant in the subjects being in acute phase of either atopic, or bacterial asthma. Moreover, IL-10 suppressed IFNγ to higher degree, than IL-4, thus favoring distinct predomination of Th2-type immune response.

Abstract. Over last years, huge efforts are made in order to improve analytic techniques for T lymphocyte subpopulations. Appropriate studies on T-helper cells are also subject to extensive research. These cellular compartments are substantially divided into various sub-classes. Such subpopulations, e.g., Th1, Th2, T-reg, Th17 and activated Т-helpers, were classified and characterized to sufficient degree. Recently, a lot of distinct data about specific receptors and functional properties of various Т-helper subpopulations was obtained. Applications of these results in laboratory research, without any doubt, will improve diagnostic quality when assessing functional alterations of immune system, as well as adequate therapy planning. (Med. Immunol., 2011, vol. 13, N 1, pp 7-16)

Аbstract. The objective of present study was to determine the immunological features of immune system aging in mucosa-associated lymphoid tissue (MALT) in the patients at different ages (mature, aging and old) observed at a dental unit. A study of cellular spectrum and humoral factors in salivary gland secretions has been performed in a group of 106 persons (35 to 90 years old). A number of age-dependent features of the immune profile were revealed for the mucous-salivary area, thus characterizing involution events within MALT structures. Among specific markers determining intensity of MALT-associated aging, a decreased percentage of viable immune cells (below 40%), along with the prevalence of the neutrophilic granulocytes in the salivary secretions (over 98%) (with increased expression of β2-integrins); decreased counts of mononuclear cells, i.e., mononuclear cells with low expression of CD11β adhesion molecules, B-lymhocytes, and Th-lymphocytes have been revealed. Alterations in serum factors included a general decrease in complement system activity (СН50) and anaphylotoxines (С3а,С5а); elevated protein, mucine, and IgМ levels. The revealed specific features of cellular and humoral immunity within MALT-associated muco-salivary zone may be considered as a normal response connected with natural aging processes. (Med. Immunol., 2011, vol. 13, N 2-3, pp 167-174)

Abstract. This review is focused on analysis of currently used flow cytometric methods designed foridentifying apoptotic cells in various invertebrate and vertebrate species. Apoptosis can be characterized by stage-specific morphological and biochemical changes that are typical to all kinds of eukaryotic cells. In this article, we consider different techniques of apoptosis detection based on assessment of cellular morphology and plasma membrane alterations, activation of intracellular enzymes and components of a caspase cascade, as well as DNA fragmentation and failure of mitochondrial transmembrane potential, as assessed in various animal groups. Apoptosis recognized as a key mechanism aiming at maintenance of cellular homeostasis in multicellular organisms, and such investigations represent a necessary component of fundamental and applied studies in diverse fields of experimental biology and immunology. A broad spectrum of apoptosis markers isused, and the preference is given to optimal approaches, as determined by experimental tasks, and technical opportunities of the laboratory.

Abstract. Flow cytometry allows estimating quantitative and qualitative structure of populations and subpopulations of immune system cells by using various methodical approaches and a wide spectrum of reagents. For diagnostics the Acquired Immune Deficiency Syndrome (AIDS) caused by a Human Immunodeficiency Virus (HIV) the flow cytometry became irreplaceable. Traditionally, immunologists examine standard model of an estimation of immune dysfunction on the basis of classical markers of Т-cells (CD3, CD4, CD8) at the HIV-infection. But researchers pay less attention to other populations and subpopulations of lymphocytes, such as γδ-, αβ- and CD38+ Т-cells. The quantitative estimation of these parameters from a HIV and AIDS patients enables to see pathogenesis a HIV infection and the prediction of its development from another side.

Abstract. Development of a modern medical science demands more thorough and deeper approach to diagnostics of diseases. New diagnostic technologies for these investigations are necessary and flow cytometry is such technology. Modern achievements in the field of fluorescent dyes, development of laser and computer technologies, have led to wide use of the given technology in medical practice. Infectious immunology is not exception. Cytokines production analysis at tuberculosis, HLA-DR expression changes for monocytes and CD64 for granulocytes at sepsis are the important diagnostic attributes. Flow cytometry offers new approaches to early and faster diagnostics of infringements of immune system at tuberculosis and sepsis, allows supervising development of these pathologies more effectively and estimate efficiency of therapy.

Abstract. Development of modern clinical and infectious immunology, demands maximum objective tests for estimation of functional condition of various types phagocytic cells, such as neutrophils, monocytes, macrophages and dendritic cells. Wide potential opportunities of flow cytometry have led to development of ways of registration of a functional condition of these cells, an estimation of processes of their activation and study of mechanisms formation of defects of their functions. For example: analysis of phagocytic and bactericidal activity of neutrophils and measurement number of the cells, which are taking place at various stages of programmed cellular death (apoptosis). Flow cytometry methods described in this review represent a new hi-tech direction for scientific and clinical researches.

We investigated the rate of biological aging, the levels of stress hormones and regulatory cytokines in Afghan war veterans and civilians with early forms of chronic brain ischemia. The study established acceleration of aging according to biological age indicators, earlier clinical onset of early forms of chronic cerebrovascular diseases associated with the processes of posttraumatic disregulation of cerebral and systemic vascular tone, the accumulation of "allostatic load” in the form of hypercortisolemia, growth regulator and angiogenic cytokines in veterans of the Afghan conflict.